The ever-increasing demand for improved productivity in research through the generation of robust analysis outputs has driven both the development and deployment of automated high-content analysis (HCA) and phenotypic cell-based approaches to drug discovery. In contrast to the more traditional cellular analysis and target-based approaches, here the researcher is able to evaluate the efficacy of potential therapeutics by monitoring the physiological state of cells through the simultaneous analysis of multiple cellular parameters in the context of an intact biological system. This course will cover the key features of HCS/A technologies and the best approaches to using these technologies for phenotypic cell-based screening.

This course has been developed to introduce and facilitate scientists who are either moving into the field or who are interested in further developing new phenotypic discovery applications and tools for use with these technologies.

Course Structure
(i) An introduction to HCA technologies
(ii) Advanced cell-based models for use with HCA
(iii) Worked examples of the phenotypic screening approach and future directions
(iv) Group discussion and Q&A

Learning Outcomes

Develop a familiarity of the basics of HCS/A technologies

Gain an understanding of the capabilities of this technology

Learn of the latest developments in cell-based models for use in this field

Get a better understanding of the key principles of assay design and development for phenotypic screening

(SC2) Exploring 3D Printing, Bioinks and Scaffolds

The promise of 3D bioprinting to create human tissues layer by layer is immense, ranging from basic biological research to drug development and testing, and ultimately to replacement organs. However, organ and tissue structures vary in complexity, and printing with living cells to create tissues is much more complicated than printing 3D objects in plastic.

Topics to be covered:

3D Printer Platforms: Inkjet vs. Pressurized Printing

3D Modeling (CAT Scans, Laser Scans and CAD)

Scaffold Selection

Cell Source Selection

Bioinks

Vascularization

This dinner course is designed for biological researchers who are interested in learning more about 3D printing and applying it to building a living tissue or organ of their choice.

What are the future medical applications in 3D printing? Can we print actual medical devices and permanent implants? Can we print drugs and living tissue? What are technical and regulatory challenges, and what’s holding us back?

There are several two-dimensional cell and biofactor printing technologies available today. Stacking and/or multilayer printing allows these technologies to be used to create three-dimensional tissue constructs. I discuss the general challenges and considerations in selecting and designing cell-ink and scaffold materials as well a method for inter-layer registration of individually addressed substrates. Detailed reference to our own Biological Laser Printing and Biopaper technologies will be used as an illustrative case study.

7:50 Multimaterial 3D Bioprinting

David Kolesky, Research Scientist, Jennifer Lewis Laboratory, School of Engineering and Applied Sciences and Wyss Institute for Biologically Inspired Engineering, Harvard University

To vastly expand the application of 3D bioprinted tissue constructs, one must be able to integrate cells, structural and vascular components concurrently and with precision. Using a custom-designed bioprinter, we are creating 3D multimaterial, cell-laden architectures with embedded vascular networks. I will describe the printing platform and our enabling ink designs as well as characterization of printed 3D cells and microstructures.

8:25 The Organovo 3D Bioprinting Platform: Changing the Shape of Medical Research and Practice

The high failure rate among clinical-stage therapies underscores the need for improved in vitro models. A common platform to produce both preclinical and therapeutic tissues would be immensely attractive. Organovo’s 3D Bioprinting platform generates microscale tissues that mimic human tissue architecture and function without exogenous biomaterial scaffolds. We will explore how medical research and practice challenges are being addressed by this novel technology platform.

Phenotypic screens of human neurons from patient-specific induced pluripotent stem cells (iPSC) can be used as discovery tools to generate new hypotheses regarding mechanisms underlying neurological disease and identify treatments based on biology rather than symptoms. An important step is to establish the technology platforms and reproducibility necessary to utilize iPSC in high-throughput drug screening. Towards this goal, we have been working to develop a standardized battery of assays against which iPSC-derived neurons can be screened for specific phenotypes. We have performed a high-content screen at the scale of 26,000 wells for chemical modulators of neurite growth using hiPSC-derived neurons, and have developed a miniaturized multiplexed high-content assay to assess neuronal ROS and mitochondrial potential. In addition, we have conducted a 5,000 well screen using Alzheimer patient-specific iPSC-derived neurons. Finally, we are developing assays to analyze neuronal function using multi-electrode arrays on multi-well plates. We will discuss our screening results and development of iPSC-based models for testing of drugs on disease-relevant cell types.

Wei Zheng, Ph.D., Group Leader, National Center for Advancing Translational Sciences, National Institutes of Health

Phenotypic screening has reemerged as an effective approach for compound screening to identify lead compounds for drug development. Recent advancement in iPS cell technology has enabled quick generation of induced pluripotent stem (iPS) cells from patient skin cells that can be further differentiated to mature cells such as neuronal cells as a disease model system for compound screening. This phenotypic disease model system is particularly useful for these diseases of which the disease pathophysiology is unclear and drug targets are not available. We have generated several iPS cell lines from patient cells with lysosomal storage diseases including Niemann Pick disease Type C, Wolman Disease and Mucopolysaccharidosis type-I. The neuronal cells differentiated from these patient iPS cells exhibit characteristic features of lipid/macromolecule accumulation and enlarged lysosomes. The results from compound screening and drug efficacy tests using these differentiated disease cells will be presented and discussed.

(SC4) Engineering Microfluidic Cell Culture Chips

Microfluidic technology holds great promise for the creation of advanced cell culture models. Engineering a microfluidic cell culture chip to emulate the dynamic physiology of a tissue’s microenvironment is paramount for primary cell culture and co-culture. As the availability of functional human cell types for in vitro culture increases, a microfluidic cell culture chip platform’s potential to produce an in vitro system capable of accurately reproducing acute and chronic human responses to drug and pathological challenges in real time will also increase.

Topics to be covered:

Cell Source Selection

Media Requirements

Channel, Chamber and Valve Design

Characterization of Microfluidic Systems

Microfabrication

Integration of Analytics

Achieving High Throughput

This dinner course is designed for researchers interested in delving into the world of microfluidic cell culture to determine its advantages over their traditional cell culture.

Detailed Agenda:

6:30 pm Welcome

6:50 Dinner Break

7:20 The Basics of Integrating Cells with Microfluidic Devices for Long-Term Cell Survival and Function in Organ-on-a-Chip Devices

Defined systems are the best option for integrating cells and tissue with microfluidic devices in controlled, reproducible assays. A defined system is composed of a known surface chemistry, serum-free medium and knowledge of cell type, differentiation state and preparation history. Examples of defined systems for cardiac, neuronal and liver as well as methods of characterization will be presented.

I will discuss various issues that are involved in creating a liver tissue model in microfluidic devices. I will also describe our recent results pertaining to a mechanism through which flow stabilizes primary hepatocyte cultures in microfluidic systems.

9:10 Interactive Q&A with Instructors and Participants

9:30 Close of Short Course

TUESDAY EVENING, NOVEMBER 18
6:00-9:00 PM

(SC5) Expert ThinkTank: How to Meet the Need for Physiologically-Relevant Assays?

It used to be adequate to build target-specific and robust assays to drive lead optimization. These assays were relatively inexpensive and reliable and could be counted on to provide chemists with usable results. However, with time, it has become apparent that it is not enough to be robust and target specific. To build therapies for patients, we need to have assays that are more predictive of patient outcome. The current buzz words are "physiologically-relevant assays." This session will explore the need for physiologically-relevant assays and explore the ways that we can achieve this endpoint.